红花水提物对乳鼠心肌细胞H2O2所致损伤的保护作用及ESR谱研究

ESR study and Protection of Water Extract of Carthamus tinctorius L. on Injury of Cardiomyocytes induced by Hydrogen Peroxide

摘要:

目的: 观察红花水提物对体外培养大鼠乳鼠心肌细胞H₂O₂损伤的保护作用及电子自旋共振(ESR)波谱的影响。 方法: 建立心肌细胞H₂O₂损伤模型,实验分5组,空白对照组,模型对照组:H₂O₂ (1 mmol·L^-1)孵育1 h;红花水提物500,100,20 mg·L^-1剂量组:分别给予不同浓度的红花水提物,预处理24 h,然后换为H₂O₂(1 mmol·L^-1)孵育1 h。实验结束后分别收集细胞上清液测定乳酸脱氢酶(LDH)、丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、一氧化氮(NO)、一氧化氮合酶(NOS)和黄嘌呤氧化酶(XOD)活性,用自旋捕集技术结合电子自旋共振波谱技术(Electron Spin Resonance,简称ESR)检测各组细胞悬液中氧自由基信号的强弱。 结果: 红花水提物各剂量组能够明显提高H₂O₂损伤后的心肌细胞的搏动频率,使细胞存活率提高,减少心肌细胞LDH的释放,降低培养液中MDA和XOD的含量,同时提高SOD,NO和GSH-Px的活性,使细胞悬液中的ESR氧自由基信号明显减弱。 结论: 红花水提物对体外培养乳鼠心肌细胞具有保护作用,其机制与其减少活性氧产生、增强氧自由基的清除、增强内源性抗氧化酶的活性有关。

Abstract:

Objective: To investigate the antioxidant effects of water extract of Carthamus tinctorius L. on injury of cardiomyocytes induced by hydrogen peroxide (H₂O₂) and to detect reactive oxygen species (ROS) for exploring the mechanisms. Methods: In cultured neonatal rat cardiomyocytes, the antioxidant effect of water extract of Carthamus tinctorius L. was investigated after H₂O₂ (1 mmol·L^-1) induced damage. The spontaneous beating, the survival rate and release of LDH, MDA, SOD, NO, NOS, GSH-Px and XOD enzyme activity in supernatant were investigated, and cell suspension was collected for detecting reactive oxygen species (ROS) by electron spin resonance (ESR). Results: Preconditioning by water extract of Carthamus tinctorius L. increased the spontaneous beating and the survival rate, and decreased the release of LDH, MDA and SOD with the rise of SOD, NO, NOS and GSH-Px enzyme activity, while in the cell suspension ROS signal decreased significantly. Conclusion: Water extract of Carthamus tinctorius L. has antioxidative effect. The mechanisms are likely related with scavenging of free radicals, enhancing its clearance, enhancing endogenous antioxidant activity.